南方医科大学学报

南方医科大学学报 ›› 2015, Vol. 35 ›› Issue (06): 832-.

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谷胱甘肽过氧化物酶2干涉慢病毒系统构建及对人肝癌细胞凋亡的影响

曹江平，唐刘君，张建宏，詹轶群，杨晓明，葛常辉   

  • 出版日期:2015-06-20 发布日期:2015-06-20

Construction a lentiviral vector for RNA interference of glutathione peroxidase 2 gene
and its effect on HepG2 cell apoptosis

  • Online:2015-06-20 Published:2015-06-20

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摘要： 目的构建人谷胱甘肽过氧化物酶2（Glutathione peroxidase 2, GPX2）慢病毒干涉载体，获得稳定下调GPX2的高滴度慢
病毒，检测敲低GPX2对细胞凋亡的影响。方法通过筛选具有显著干涉效果的siRNA序列，构建pSicoR-GPX2慢病毒干涉载
体，包装病毒并感染人肝癌HepG2 细胞，分别用Western-blot 和RT-PCR方法检测GPX2 表达情况，应用流式细胞术分析敲低
GPX2对人肝癌细胞凋亡的影响。结果成功构建pSicoR-GPX2干涉慢病毒载体并获得高滴度慢病毒颗粒，GPX2蛋白表达水
平和RNA表达水平均有显著下调，且人肝癌细胞感染GPX2干涉慢病毒后对凋亡更加敏感，其原因可能是促凋亡蛋白Bax的活
化和抗凋亡蛋白Bcl-2活性的抑制。结论成功构建人GPX2基因干涉慢病毒，为肝癌靶标治疗提供新的线索，为后续GPX2功
能研究奠定基础。

Abstract: Objective To construct a RNA interference lentiviral vector for human glutathione peroxidase 2 (GPX2) gene and
observe the effect of GPX2 knockdown on cell apoptosis. Methods The sequence of the small interfering RNA (siRNA) for
GPX2 interference was inserted into the pSicoR vector. HepG2 cells were infected by the packaged si-GPX2 lentivirus and the
expression of GPX2 in the infected cells was detected by both RT-PCR and Western blotting. Changes of cell apoptosis
following the infection were analyzed by flow cytometry. Results The lentiviral particles pSicoR-GPX2 were successfully
packaged. The expression of GPX2 in the infected cells was obviously down-regulated at both RNA and protein levels. GPX2
knockdown caused increased apoptosis rate, increased Bax expression and lowered Bcl-2 expression in HepG2 cells.
Conclusion We have successfully constructed the lentiviral vector for RNA interference of human GPX2 gene.